UPLC-MS/MS法检测动物性食品中19种β-受体激动剂残留

Determination of β-agonists in Animal Derived Food by UPLC-MS/MS

建立了前处理过程较为简便、能同时检测猪、牛和羊的肌肉、肝脏、肾脏和鸡蛋中特布他林、齐帕特罗、沙丁胺醇、西马特罗、西布特罗、克仑塞罗、羟甲基克伦特罗、克仑丙罗、氯丙那林、莱克多巴胺、克仑特罗、妥布特罗、溴布特罗、克仑潘特、班布特罗、马布特罗、马喷特罗、苯乙醇胺A和喷布特罗19种β-受体激动剂的UPLC-MS/MS方法。样品经乙腈-异丙醇混合溶液（4∶1,V/V）提取后,用β-葡萄糖醛苷酶/芳基硫酸酯酶酶解,混合型阳离子交换固相萃取柱净化,然后用UPLC BEH C18色谱柱分离,以0.1%甲酸乙腈溶液和0.1%甲酸水溶液为流动相进行梯度洗脱,基质匹配溶液内标法或外标法定量。结果表明：19种β-受体激动剂在1~50 ng/m L基质匹配标准溶液浓度范围内呈现良好线性关系,相关系数R2均大于0.990;在猪、牛和羊的肌肉、肝脏、肾脏和鸡蛋中的检测限均为0.1 ng/kg,定量限均为0.5 ng/kg。从0.5、1、2和5 ng/kg四个添加浓度检测结果可以看出,19种药物的回收率范围为60%~120%,批内、批间相对标准偏差均小于20%。该方法具有简便快捷、灵敏度高、定性准确等特点。

A UPLC-MS/MS method was established for the determination of terbutaline,zilpaterol,salbutamol,cimaterol,cimbuterol,clenciterol,hydroxymethyl clenbuterol clenproperol clorprenaline,ractopamine,clenbuterol,tulobuterol,brombuterol,clenisopenterol,bambuterol,mabuterol,mapenterol,phenyl ethyl alcohol amine A and penbutolol residue in animal derived food. After the tissues were extracted by Acetonitrile-2-Methyl-1-propanol,enzymolysis,centrifuged,neutralized,followed by liquid-liquid extraction with ethyl acetate and tert-butyl methyl ether,respectively. The combined extracts were applied to a solid phase extraction MCX cartridge for cleanup. The separation of β-agonists was performed on Waters Acquity UPLC system with the column of BEH C18 and the gradient elutinon solvent of acetonitrile（ 0. 1% formic acid） and water（ 0. 1% formic acid） at a flow rate of0. 3m L / min. The method was quantified by blank tissue spiked standard curves. The calibration curves were good linear between the peak areas and the concentrations of 1 ~ 50μg / kg,the correlation coefficient R2〉 0. 990. The limits of detection of the 19 β-agonists in animal derived food were 0. 1μg / kg,and the limit of quantification was0. 5 μg / kg. The average recoveries from spiked animal tissues at three concentrations of 0. 5,1,2 and 5 μg / kg ranged 60% ~ 120%. The intra-and inter-batch variation coefficients were both less than 20%.